HIF-1α survivin在宫颈上皮内瘤变及宫颈鳞癌中的表达和临床意义

目的：研究缺氧诱导因子-1α（HIF-1α）和生存素蛋白（survivin）在正常宫颈组织、宫颈上皮内瘤变组织（CIN）及宫颈鳞癌组织中的表达及意义。方法应用免疫组化的方法检测HIF-1α、survivin在宫颈鳞癌标本74例、CIN标本40例、正常宫颈标本24例中的表达情况。分析了两者与宫颈鳞癌病理学分级、临床分期及淋巴结转移的关系。结果 HIF-1α、survivin两种蛋白在正常宫颈组织中不表达,但在CIN组和宫颈鳞癌组中阳性表达率逐渐升高;HIF-1α、survivin阳性表达率与宫颈鳞状细胞癌的病理分级,临床分期,淋巴结转移呈正相关;在宫颈癌组织中HIF-1α与survivin的表达程度呈正相关。结论缺氧诱导因子-1α与survivin的阳性表达对恶性肿瘤的发展起着重要作用,对两个指标的联合检测有助于评估肿瘤的恶性程度及判断预后。     

原花青素对人骨肉瘤Saos-2细胞增殖的影响及机制研究

目的研究原花青素体外对人骨肉瘤Saos-2细胞增殖的影响及其作用机制。方法体外培养人骨肉瘤Saos-2细胞,MTT法检测细胞增殖抑制率;PI染色法检测细胞凋亡率;ELISA法检测Caspase-9、Caspase-12、Survivin含量;Bradford法检测SOD、M DA含量。结果 PC在48 h和72 h对Saos-2细胞的体外增殖均具有抑制作用,并具有浓度和时间依赖性;40、80、160μg/m L浓度的PC均诱导Saos-2细胞凋亡;促进Caspase-9和Caspase-12蛋白表达,抑制Survivin蛋白表达;增加细胞内SOD的活性,降低M DA的含量。结论 PC可抑制Saos-2细胞的增殖,可能与其增加机体的抗氧化功能及启动死亡受体通路、内质网通路、线粒体通路这三条途径诱导细胞凋亡有关。     

Adenovirus expressing p27kip1 suppresses growth of established esophageal carcinoma xenograffs

AIM: To investigate the growth suppression of adenovirus expressing p27kip1 on established esophageal tumors in nude mice.METHODS: Esophageal carcinoma xenografts in nude mice were established by tumor tissue mass transplantation. The successfully constructed recombinant adenoviral vectors carrying p27kip1 gene (Adp27kip1) were directly injected into the esophageal tumors in nude mice. Compared to control group, the growth curve of tumor was drawn and the growth inhibition rate of tumor was calculated. The histology of tumors was examined by hematoxylin and eosin (H&E) staining. The expression of p27kip1 and survivin was detected in tumors by immunohistochemical technique.RESULTS: The growth of tumors in gene therapy group with Ad-p27kip1 was obviously suppressed compared to control group (0.42±0.08 g vs 1.17±0.30 g, t=6.39,P＜0.01), the inhibition rate of tumor growth reached 64.1%. Pathological detection showed that the tumors in nude mice were poorly differentiated esophageal squamous carcinoma. In addition, the expression of p27kip1 was increased, while the expression of survivin was decreased in tumors after being transfected with Ad-p27kip1.CONCLUSION: p27kip1 gene therapy mediated by adenovirus vector has a significant inhibitory effect on esophageal carcinoma in vivo. Up-regulated p27kip1expression and down-regulated survivin expression may be its important mechanisms.     

Survivin与VEGF在肺癌中的表达及意义

目的探讨Survivin及血管内皮生长因子(VEGF)在肺癌组织中的表达及其与患者预后的关系方法应用免疫组化法检测化疗组和非化疗组肺癌患者Survivin、VEGF的表达水平,比较Survivin、VEGF的表达与生存宰的关系。结果Survivin和VEOF在挪癌组织中的阳性表达率分别为74．3％和70．0%,二者呈正相关(r=0．562,P<0．01);两组Survivin和VEOF阴性表达者生存期明显长于阳性表达者。结论Survivin VEGF在肺癌中的表达呈正相关;二者联合检测是患者化疗疗效、预后判断的重要指标。     

Survivin反义寡核苷酸抑制肝癌移植瘤生长的作用

目的:检测Survivin反义寡核苷酸(ASODN)在人肝癌耐药细胞株裸鼠皮下移植瘤中的表达情况．方法:将30只裸鼠建立人肝癌耐药细胞系 SMMC-7721／ADM皮下移植瘤模型,随机分成6组:空白对照组(A)、脂质体转染对照组 (B)、正义链对照组(C)、200(D)、400(E)和 600 μg/L(F)反义链组(ASODN组),用不同的转染液注射后2,4,8,12,16,20 d,用逆转录聚合酶链反应技术(RT-PCR)和Western blot蛋白免疫印迹法检测治疗后各组肿瘤组织中Survivin mRNA和蛋白表达的变化．结果:注射后20 d,ASODN组肿瘤细胞生长明显抑制,空白对照组、脂质体转染对照组和正义链对照组裸鼠的mRNA和蛋白表达无明显差异,而ASODN组mRNA和蛋白表达随着时间和浓度的增加,Survivin表达减弱,E,F组与其余个组(A,B,C,D)相比有显著差异(mRNA: 0．33±0．04,0．28±0．03 vs 0．82±0．02,0．78± 0．05,0．72±0．04,0．57±0．03,P<0．05;蛋白: 34．9±3．89,21．2±3．65 vs 72．14±6．53,69．31 ±5.34,68．29±4．98,53．8±5．23,P<0．05)．结论:Survivin反义寡核苷酸能够下调 Survivin mRNA和蛋白的表达,抑制裸鼠皮下移植瘤的生长．     

纤维支气管镜活检病变组织标本和痰标本Survivin基因的检测对肺癌的诊断价值

目的明确纤维支气管镜(以下简称纤支镜)活检病变组织和痰标本中SurvivinmRNA的检测在肺癌诊断中的意义。方法应用逆转录聚合酶链反应(RT PCR)法检测41例肺癌手术标本癌组织、癌旁组织和9例良性肺疾病病变组织手术标本,80例肺癌和30例良性肺疾病纤支镜活检病变组织标本及所有(160例)患者痰标本SurvivinmRNA表达情况,并与病理组织学、刷检细胞学和痰细胞学检查结果比较。结果肺癌手术切除标本癌组织SurvivinmRNA的阳性率为70.7%(29/41),高于癌旁组织[17.1%(7/41)]和良性肺疾病组织(1/9),差异有统计学意义(χ2值分别为23.97和10.93,P均<0.05),而癌旁组织与肺良性疾病组织相比,差异无统计学意义(χ2=0.20,P>0.05);纤支镜活检肺癌组织标本SurvivinmRNA的阳性率为63.8%(51/80),高于良性肺疾病的13.3%(4/30,χ2值为22.18,P<0.05);肺癌患者癌组织SurvivinmRNA表达与否及表达水平与患者年龄、性别及肿瘤的病理分型、分级、部位及转移情况均无明显相关性(P均>0.05)。肺癌患者痰标本SurvivinmRNA的阳性率是59.5%(72/121),癌细胞的检出率是47.1%(57/121);痰Survivin mRNA检测联合痰细胞学检查诊断肺癌的敏感性为80.2%(97/121),高于单独痰细胞学及单独痰SurvivinmRNA检测的敏感性(P均<0.05)。手术标本、纤支镜活检标     

Expression of a novel apoptosis inhibitor-survivin in colorectal carcinoma

AIM: To investigate the role of survivin expression in the pathogenesis of colorectal carcinoma. METHODS: Immunohistochemistry S-P method and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were used to detect the expression of survivin and apoptotic cell in situ in colorectal cancerous tissues, para-cancerous tissues and normal tissues of 48 cases of colorectal carcinoma. RESULTS: The survivin positive unit (PU) was higher in cancerous tissues (38.76±5.14) than in para-cancerous (25.17±7.26) or normal tissues (0.57±0.03) (P<0.05). The apoptosis index (AI) of para-cancerous tissues was (7.51±2.63%) higher than cancerous tissues (4.65±1.76%). The expression of survivin was associated with pathological grade, lymph node metastasis and Dukes stage of colorectal carcinoma. CONCLUSION: Survivin expression may play an important role in carcinogenesis of colorectal carcinoma and may be associated with malignant biological behaviors of colorectal carcinoma.     

Transgenic expression of survivin compensates for OX40‐deficiency in driving Th2 development and allergic inflammation

Survivin, an inhibitor of apoptosis family molecule, has been proposed as a crucial intermediate in the signaling pathways leading to T‐cell development, proliferation, and expansion. However, the importance of survivin to T‐cell‐driven inflammatory responses has not been demonstrated. Here, we show that survivin transgenic mice exhibit an increased antigen‐driven Th2 lung inflammation and that constitutive expression of survivin reversed the defective lung inflammation even in the absence of OX40 costimulation. We found that OX40‐deficient mice were compromised in generating Th2 cells, airway eosinophilia, and IgE responses. In contrast, OX40‐deficient/survivin transgenic mice generated normal Th2 responses and exhibited strong lung inflammation. These results suggest that OX40 costimulation crucially engages survivin during antigen‐mediated Th2 responses. These findings also promote the notion that OX40 costimulation regulates allergic responses or lung inflammation by targeting survivin thereby enhancing T‐cell proliferation and resulting in more differentiated Th2 cells in the allergic inflammatory response.     

Functional Characterization of Novel Mutations Affecting Survivin (BIRC5)‐Mediated Therapy Resistance in Head and Neck Cancer Patients

Survivin (BIRC5) is an acknowledged cancer therapy‐resistance factor and overexpressed in head and neck squamous cell carcinomas (HNSCC). Driven by its nuclear export signal (NES), Survivin shuttles between the nucleus and the cytoplasm, and is detectable in both cellular compartments in tumor biopsies. Although predominantly nuclear Survivin is considered a favorable prognostic disease marker for HNSCC patients, the underlying molecular mechanisms are not resolved. Hence, we performed immunohistochemical and mutational analyses using laser capture microdissection on HNSCC biopsies from patients displaying high levels of nuclear Survivin. We found somatic BIRC5 mutations, c.278T>C (p.Phe93Ser), c.292C>T (p.Leu98Phe), and c.288A>G (silent), in tumor cells, but not in corresponding normal tissues. Comprehensive functional characterization of the Survivin mutants by ectopic expression and microinjection experiments revealed that p.Phe93Ser, but not p.Leu98Phe inactivated Survivin's NES, resulted in a predominantly nuclear protein, and attenuated Survivin's dual cytoprotective activity against chemoradiation‐induced apoptosis. Notably, in xenotransplantation studies, HNSCC cells containing the p.Phe93Ser mutation responded significantly better to cisplatin‐based chemotherapy. Collectively, our results underline the disease relevance of Survivin's nucleocytoplasmic transport, and provide first evidence that genetic inactivation of Survivin's NES may account for predominantly nuclear Survivin and increased therapy response in cancer patients.